Int J Anat Res 2015, 3(1):922-26. ISSN 2321-4287
Original Article
INCAPABILITY OF FORM ALIN BASED FIXATIVE TO KILL M AGGOT
IN M UTILATED HUM AN CARRION
Oladele, A.A. *
1, Sanya, J.O.
2, Raji, M .O
3, Ekundina, V.O
3.
ABSTRACT
Address for Correspondence: Dr. Oladele, A.A., Depart m ent Of Hist opat hology And M orbid Anat omy, Obafem i Aw olow o Universit y Teaching Hospit al, Ile-Ife, Osun State, Nigeria. Call No.: +2348088176682 E-M ail: ladelerot i45@gm ail.com
* 1 Depart m ent Of Hist opat hology And M orbid Anat omy, Obafem i Aw olow o Universit y Teaching
Hospit al, Ile-Ife, Osun St at e, Nigeria.
2 Depart m ent of Physiological Science, Afe Babalola Universit y, Ado-Ekit i, Nigeria.
3 Depart m ent of M edical Laborat ory Science, Afe Babalola Universit y, Ado-Ekit i, Nigeria.
Background:Em balm ing is a pr ocess used t o t em por arily preserve a hum an cadaver t o forest all decom posit ion and m ake it suit able f or display at f uner als; t hus, are agent s t hat pr event aut olysis and put r efact ion. The out break of m aggot s fr om a heap of inadequat ely em balm ed bodies due t o deep cut s and bodies involved in inf erno, necessit at ed t he need t o re-invest igat e t he efficacy of for m alin based em balm ing fluid and it s inabilit y t o kill m aggot s.
M et hodology: Var ious st rengt h of Form aldehyde, Xylene, Kerosene, and, Lim e fluid, Isopropanol, Gam alin 20, Pot assium ferrocyanide, and Physiological saline as cont rol w ere used in t he invest igat ion. In t he present invest igat ion, Tw o m aggot s under t he sam e at m ospher ic condit ion w ere put in each of t he t en select ed chem ical reagent s/ solut ions, including Lim e, Kerosene, and t he Gam alin 20 t hat are nat urally available w ere init ially dispensed int o t en glass universal cont ainers. M aggot m ovem ent s in each reagent solut ion w ere cr it ically observed.
Result: M aggot s deat h occurred w it hin t he first t en m inut es in t est num ber t hr ee gr oups III t hat cont ains Concent rat ed Form alin and Xylene and M aggot s died aft er fift een m inute of the experim ent , but m aggot s did not died unt il about eight hour s aft er t he t est in t w o of t he experim ent .
Discussion:Result s of t his invest igat ion show ed clearly t hat M aggot s w er e not killed as soon as expect ed by t he em balm er w hen ordinary t en percent alcoholic form alin em balm ing fluid is used. Equal volum e of concent rat ed for m alin plus Xylene w as found out t o be effect ive at killing m aggot instant ly.
Conclusion: It is t herefore advisable t o use Xylene plus Conc. Form alin w hen preserving cadaver infest ed w it h M aggot and t his could at t he sam e t im e pr event t he occurrence of M aggot s Infest at ion and bet t er pr eservat ion of m ass of burnt m ut i lat ed cor pses in our M ort uary.
KEY W ORDS:Em balm ing, Fixat ives, M aggot , Hum an Carr ion, Xylene.
INTRODUCTION
Int ernat ional Journal of Ana tomy and Research, Int J Anat Res 2015, Vol 3(1):922-26. ISSN 2321- 4287 DOI: ht t p:/ / dx.doi.org/10.16965/ ijar.2015.117
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Received: 10 Feb 2015 Accept ed: 16 M ar 2015 Peer Review : 10 Feb 2015 Published (O):31 M ar 2015 Revised: 26 Feb 2015 Published (P):31 M ar 2015
Int ernat ional Journal of Anat om y and Research ISSN 2321-4287
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DOI: 10.16965/ ijar.2015.117
M aggot is a larva of t he fly of order Dipt era is p ar t i cu l ar l y r el at ed t o Br ach ycer an f li es exam ples of w hich are cheese flies, green bot t le flies, blow flies, and house flies. This is unlike
M ost flies are not very friendly to human because of t heir process of t ransm it t ing large num ber of com m unicable diseases, and product ion of filthy m aggot by m ost of t he flying insect s. They are com monly found in filthy and dirt y environment . Accordingly t hey are alw ays signals t hat indicat e t he presence of dead decaying bodies around. This spans t hrough insect s, and anim als. Alm ost all t he gravid fem ale adult flies belonging t o Green bot t le fly species, m et aphozise from egg laying t o M aggot s (larvae) product ion. They alm ost appear as cream y-w hit e st ruct ure and conico-cylindr ically in shape. Essent ially in average t hey ar e about 15 – 20m m lengt h [ 2,3,4] . M aggot s ar e usu all y seen o n decom posing m at t er such as garbage and on dead m ult icellular organism s especially on dead birds, anim als, and hum an organism , t herefore serve as an essent ial com ponent of t he eco-syst em [5].
Em balm ing is a w ay of t em porarily dist urbing t he ecosyst em by preserving a hum an cadaver t o forestall decom posit ion and m ake it suitable for display at a funeral; and, Post M ort em , and Forensic st udy in crim e invest igat ion. This is a form of pre- t reat m ent of corpses w it h special preser vat ives m ainly chem icals in or der t o prevent decom posit ion. Perfum e or fragrance, even eyelashes and false eyes are added t o t he bodies w hen it com es t o funeral present at ion. Thus, agent s r esponsible f or aut olysis and put refact ion are alm ost halt ed. This event ually prevent ed maggot infest ation, because noxious sm elling debris produced by put refying Bact eria and aut olysis had been st opped .How ever, whenever t he opposite happened then there will be m aggot infest at ion as long as flies are not prevented from landing on t he dead m atter. Form aldehyde, Alcohol, Phenol, Glycerol and w at er are t he m ajor chem ical agent s used in m ost em balm ing technique t oday, except in t he case of Thiel em balm ing t echnique w hich is based on 4-chlor o 3- m et hylenephenol f or fixat ion, boric acid for disinfect ing and et hylene glycol for preservat ion of t issue plast icit y [6]. Form alin is an organic com pound w it h form ulae CH2O. It is a sim plest form of aldehyde. It is w idely used as it is alw ays readily available [7, 8].
Form alin fixes t issues by form ing a m et hyl cross
linkages w it h prot ein t hereby replacing t he norm al colloidal fluid in t he cells w it h a jelly-like rigid com pound. The latter effect exhibit s t he coagulat ion propert ies of form aldehyde. Ti ssu e an d b act er i um cel l s ar e m ad e o f prot oplasm and as such, cont ain large am ount s of m oist ure. The int roduct ion of form aldehyde int o t h e t i ssue coagulat es pr ot eins in t he prot oplasm living am bient water, t he expected dryness t hat m ay follow init ial em balm ing is norm ally arrest ed by inject ing t he corpse w it h addit ional t w o percent (2%) form alin and t his process killed t he cell. In t erm s of em balm ing pract ices, t his is a perf ect sit uat ion as t he form aldehyde not only disinfect s t he t issue but replaces t he t issue cell w at er w it h a rigid gel t hus allow ing t he em balm ed t issue t o m aint ain its cont our. Addit ionally, t he “ new ” cell st ruct ure w il l r esi st f u r t h er b act er i al at t acks as it s com posit ion now cont ains a f orm aldehyde-based com pound w it h t he t issue elem ent s t hereby denat ure t he t issue. Form aldehyde is a pot ent disinfect ant and ant i-bact erial agent [9,10] and used for fum igat ion(s), less effect ive as a fungicide, insect icide, or larvicide [9]. Form alin fixes t issue very slow ly w hen compared w ith alcohol and acet ic acid but penetrat e t issue very fast [11]. A t issue of 4m m in t hickness w ill be w ell penet rated in less t han one hour. This r elat es t o t h e bi nd ing t im e necessar y f or form aldehyde t o fix a t issue, a 90% t hreshold binding takes not less t han 24 hours at 22C and 18hours at 37C respect ively. The t issue does not die unt il it is fixed and t he process is reversible if w it hdraw n before t he cell deat h (s) [8]. The rat e of penet rat ion of form alin is said t o be d = k x square root of t im e, w here d is t he dept h of penet rat ion, k is t he coefficient of diffusion w hich is bet w een 2.0 – 3.6 for concent rat ed form alin, 0.78 for 10% form alin [12]. The first layer is penet rated at the rate of 90mm per hour. The 4m m t issue block w ould be fully penetrat ed in less t han 1 hour [13].
Int J Anat Res 2015, 3(1):922-26. ISSN 2321-4287 Recent ly w e found out t hat M aggot breakout from a m ass of bodies t hat w ere burnt beyond recognit ion from a ghast ly road Traffic Accident (RTA) t hat result ed int o a huge inferno. The b o di es w er e b r ou gh t t o t h e ou r Ho sp it al M ort uary and w ere t reat ed as usual by form alin based fixat ive hoping t hat the bodies w ill rem ain int act t ill som e days prior t heir release t o relat ives t o be taken away for burial in t his case m ass burial is usually done. But t o our dism ay w e found out t hat t he bodies have been infested w it h num erous num ber of flies m aggot s, aft er em balm ing t hem and put t hem in an enclosed room w it h air condit ioner as pract iced. This is w hat act ually encourages us t o carry out t he present invest igat ion. This idea now suggest s t hat m aggot s w ere likely presented t hat t hey have capabilit y t o resist t he f or m alin base fixat ion.
In view of t he above curiosit y, som e chem icals carefully select ed in our Laborat ory and w ere put t o t est . One of t he select ed chem ical m ixt ures w as Physiological saline w hich w as used as negat ive cont rol and w e found out t hat of som e t he chem ical t est ed against living m aggot s w ere killed im m ediat ely they were put int o cert ain m ixt ure, w hile it t ook som e days b ef o r e m aggo t s pl aced i n so m e o t h er chem icals w ere killed. Our observat ion show s t hat som e of t he m ixt ures w ere very effect ive at k il l in g t h e M aggot s w h il e i n so m e t h e chem icals are found out t o be poorly effect ive at killing m aggot s placed in such m ixt ure. The present invest igat ion discovered som e chem ical m ixt ures t hat kill M aggot s at a fast er rat e t han ot hers in t he groups invest igat ed.
M ATERIALS AND M ETHODS
Form alin fixed and inject ed M ass of hum an carrion but st ill infest ed wit h num erous m aggot s w as used in obt aining act ively living m aggot s for t he experiment . Glove hand was used t o pick t he m aggot s. This w as t r ansferred int o t he laborat ory very close t o t he M orgue. In t he laborat ory four m aggot each w ere put int o each of t he solut ion / m ixt ure int o each of t he 10 universal bot t les using a long blunt end forceps t o picked m aggot and dropped int o t he universal bot t les.
RESULTS
Four m aggot s each of bet w een (8-16m m in lengt h) from t he heap or pile of burnt hum an bodies aft er em balm ing w ere put int o each of t he follow ing solut ions: - Physiological saline, 10% form al saline, Concent rat ed form alin plus Xylene (Xylol), Concent rated form alin only, Xylol on ly, Gam al in 20, Lim e p lu s con cent r at ed form alin, Kerosene plus concent rat ed form alin, Sat urat ed solut ion of Pot assium ferrocyanide, and finally Concentrat e Isopropanol. All m ixture/ Solut ions w ere dispensed in 30m l equal volum e int o glass un iver sal bot t les ar r anged on a laborat ory w orking bench in a fully illum inat ed laboratory for easy and correct observat ion. The t im e it t ook for t he m aggot s t o die in each of t he t est and cont rols w ere carefully observed and recorded im m ediat ely (Dat ed 05-10-2014, 9:42am – 6:00pm .
The m aggot s in group III (Conc. Form alin + Xylene) died w it hin t he first 10m inut es, but t hose m aggot s in groups II, (10% Form alin), VII (Lim e + Conc. Form alin, and VIII (Kerosene) died at 15m inut es int erval ,how ever , m aggot s in group IV (Conc. Form alin) died in t he next 20m inut es intervals aft er t he com m encement of t he experiment..
M aggot s in Groups I (Physiological saline),
V (Xylene + 10% Form alin), and VI (Gam alin 20), w ere deadly sluggish at 40 m inut es int erval, IX (Sat ur at ed Pot assium Fer r ocyanide) and X (Isop r o p an ol ) gr o up s w er e sl uggi sh at 40 m inut es at t he sam e t im e.
The m aggot in groups I, V, and VI were still deadly sluggish even aft er 3hours post experim ent , but finally died at 8hours int erval post experim ent . The m aggot in groups XI and X t hough w ere sluggish at t hree hour, but finally died at 8t h
hour int erval of t he experim ent .
Act ivit ies of som e chem icals on m aggot s in t he hist opat hology laborat ory.
Te st / t im e 0-5 m ins 5-10 m ins 10 -15 m ins 1 5-20 mins 2 0-25 m ins 25 -30 m ins 3 0-3 5 m ins 35 -40 m ins
₊ ₊ ₊
II 1 0% Form alin ₊₊₊ ₊₊ ‡ ‡ ‡ ‡
I Physiological saline ₊₊₊ ₊₊₊ ₊₊ ₊₊ ₊
‡ ‡
III Concent rat e d Formalin+ Xylol ₊₊₊ ‡ ‡ ‡ ‡ ‡ ‡ ‡
IV Formalin only ₊₊₊ ₊₊₊ ₊ ‡ ‡ ‡ ‡ ‡
₊ ₊ ₊
VI Gam alin 2 0 ₊₊₊ ₊₊₊ ₊₊ ₊₊ ₊₊ ₊
V Xylol only ₊₊₊ ₊₊ ₊₊ ₊₊ ₊
₊ ₊
VII Lim e juice+Conc. Form alin ₊₊₊ ₊₊₊ ‡ ‡ ‡ ‡ ‡ ‡
VIII Ke rosene+ Formalin ₊₊₊ ₊₊ ‡ ‡ ‡ ‡ ‡ ‡
₊₊₊ ₊₊₊
IX Sat urat e d Pot . Fe rrocyanide ₊₊₊ ₊₊₊ ₊₊₊ ₊₊₊ ₊₊₊
X Isopropanol ₊₊₊ ₊₊₊ ₊₊₊ ₊₊₊ ₊₊₊ ₊₊
₊₊₊ ₊₊₊ ₊₊
ACTIVE M OVEM ENT +++ , SLUGGISH M OVEM ENT ++ , DEADLY SLUGGISH M OVEM ENT + , DEAD (ZERO M OVEM ENT ‡
DISCUSSION AND CONCLUSION
Xylene, Kerosene, and form al- Lim e w ere found t o be very act ively t oxic t o m aggot , t he four m aggot s each died w it hin first 3- 5m inut es. Xylene, kerosene, and Lim e form ol had not been used. The m aggot s w ere able t o live longer in 10% form alin, equal volum e of form alin and alcohol for 1hr 30m in and 1hr respect ively. Kerosene and Xylene proved t o be m ore let hal to m aggots as both m ixtures cont aining Formalin killed m aggot s w it hin 5 m inut es. The longer period t hat m aggot s st ayed alive in form alin support t he fact s t hat form alin is less effect ive at dest roying M aggot s or Grubs of m ost fly larvae.[9]
Form alin only penet rat es t issue very fast but fixes slow ly [12]. And deat h only occurs w hen a t issue is fixed. M aggot s used in t his experiment were about 15mm in length and 4mm in widt h. This signified t hat formalin penet rat ion w ould be achieved in less t han 30 m inut es, calculat ing from t he form ula d = k x square root of t im e, d= 4m m w idt h of t he m aggot , const ant k is t aking t o be 0.78. Fixat ion w hich is t he form at ion of m et hyl cross linkage convert ing t he t issue const it uent t o irreversible gel m ay t akes a lot of t im e aft er penet rat ion as 90%
t hreshold binding m ay take 24 hours at 22o
C and 18 hours at 37oC. This vindicat es t he abilit y of m aggot t o live f o r such a long t im e w hen im m ersed in a form alin solut ion.
We decided t o use Xylene w it h equal volum e of concent rat ed formalin is because of t he fact s t hat using Xylene alone m ay burn t he t issues
rather t han preserving t he tissues, w hich could even burn a hum an skin if com e in cont act . It has never been used as part s or com ponent of em balm ing fluid. t o It s our t hinking t hat xylene m ay act as a form of Fungicide and t hat inabilit y of form alin at killing M aggot m ight be that Grubs t issue cont ain cert ain subst ance t hat prevent form aldehyde penet rat ion . We hope t hat fut ure researchers can a furt her st udy on t his problem . Th e f ast est d eat h of m aggo t o ccur r ed i n so lu t i on / m ixt u r e o f f or m ali n pl us Xyl en e, Kerosene and lim e respect ively. In tandem w it h current invest igat ion it is pert inent t o use any of Xylene, Kerosene or Lim e juice w henever out br eak of Gr ub inf est at ion is su spect ed especially bodies brought int o the M ort uary due road t raffic accident .
The em balm ing fluid for hum an corpses w it h open w ound should cont ain m ore phenol w hen t ot al im m ersion is not possible.
Conflicts of Interests: None
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